I try a new method for chloroplasts analysis and its my first experiment with gradient electrophoresis for membrane proteins. In the protocol is Bis-Tris used, in lab we have Tris-HCl, the working pH range is 7-7,6, and the running buffer is MES. So, especially, can be used Tris/HCl and MES together and can be Bis-Tris in this systen already replaced? Thanks :)
Hi there,
At the working pH Bis-Tris molecule would be vastly deprotonated (as its pK is 6.5) and neutral whereas Tris molecule would be protonated (pK 8.1) and positively charged. So chemically speaking they are not equivalent.
You should first get some info about buffers and where the molecules really buffer.
Hi Tereza, I would stay away from Tris-HCl because it is an acid salt of tris. You need to add NaOH to the tris-HCl to bring it up to the right pH. The added salt will ruin any gel. I made that mistake once thinking that tris-HCl and tris free base were the same. They are not.
You should work with Tris free base when doing electrophoresis..
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biochemical Techniques
- Analytical Chemistry
- pH